Development of a PCR assay combined with a short enrichment culture for detection of Campylobacter jejuni in estuarine surface waters

• Published in: FEMS microbiology letters Vol. 127; no. 3; pp. 201 - 206
• Main Authors: Hernandez, J, Alonso, J.L, Fayos, A, Amoros, I, Owen, R.J
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.04.1995; Blackwell
• Subjects: bacterial proteins; Bacteriological methods and techniques used in bacteriology; Bacteriological Techniques; Bacteriology; Base Sequence; Biological and medical sciences; Brackish; Campylobacter jejuni; Campylobacter jejuni - genetics; Campylobacter jejuni - isolation & purification; cell culture; detection; DNA; DNA Primers - genetics; DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; extraction; flagellin; Fundamental and applied biological sciences. Psychology; Microbiology; Molecular Sequence Data; Polymerase chain reaction; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - statistics & numerical data; Seawater; Sensitivity and Specificity; Surface water; Water Microbiology; Culture medium; Selective medium; Polymerase chain reaction; Brackish water environment; Campylobacteraceae; DNA; Surface water; Bacteria; Method; Detection; Campylobacter jejuni
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.1995.tb07474.x

Two extraction procedures were examined, and it was found that DNA recovered from Campylobacter jejuni lysed by the cetyltrimethylammonium bromide (CTAB) method was more suitable for use as a PCR template than DNA released by the boiling method. The region targeted for PCR amplification was a 1.73-kb portion of the flagellin A gene of C. jejuni. The detection limit was lower than 30 cells per 100 ml in artificially contaminated waters. PCR assay and conventional culturing method had the same sensitivity, but results of the PCR technique were available within 48 h and so shortened the time necessary for detection by 48 h.